| Background Acute myeloid leukemia(AML)is the most common type of acute leukemia,mainly due to the abnormal proliferation of large numbers of primitive cells in the bone marrow.Traditional first-line treatment for AML is mainly based on chemotherapy,but the survival curve of patients is stagnant.The search for targeted drugs to improve remission rates,survival and quality of life in AML patients is currently a hot topic in AML research.Venetoclax,a competitive inhibitor of BCL-2,has a key role in tumor cell survival and treatment resistance in AML,but its single-agent activity is limited,and a therapeutic regimen based on "Venetoclax + X" is currently a hot topic for clinical exploration.Proteasome inhibitors are more sensitive to the effects of leukemic cell action.Ixazomib,a new generation proteasome inhibitor,has received much attention in studies for the treatment of AML.Clinical studies have found that Ixazomib significantly improves remission rates in combination therapy.Therefore,we investigated the combination of vincristine with Ixazomib in the treatment of AML to see whether the two drugs can synergistically promote apoptosis and inhibit cell proliferation and to further investigate the mechanism of action.Objective We studied AML cell lines and primary cells of AML patients to clarify the effects of Venetoclax combined with Ixazomib on AML cell viability and apoptosis,and to explore the molecular mechanism of the synergistic enhancement of AML cell killing by the combination of the two drugs.By building an animal model to analyze the degree of tumor cell infiltration in bone marrow,the synergistic inhibitory effect of the two drugs was further verified,providing a research basis for new treatment options for AML.Methods(1)In vitro experiments were performed on AML cell lines and primary cells to investigate the effects of Venetoclax combined with Ixazomib on cell viability and apoptosis levels.Cell viability was measured by Cell Titer-Lumi,and the synergistic effect was calculated by Compusyn;the apoptotic ratio was measured by flow cytometry;the expression level of relevant apoptotic proteins was detected by WB;the expression of differential genes after drug treatment was analyzed by transcriptome sequencing and pathway enrichment was performed and verified by WB;(2)In vivo experiments were performed in NSG mice.MOLM-13-Luciferase cells were used to construct a CDX animal model,and the mice were randomly divided into four groups and then treated with drugs.Results(1)Venetoclax and Ixazomib alone were cytotoxic in AML cell lines in a dose-dependent manner;(2)Venetoclax combined with Ixazomib significantly inhibited the proliferation of AML cell lines and primary cells of AML patients and upregulated the expression level of apoptotic proteins,indicating that the combination of the two drugs could promote tumor cell apoptosis;(3)In vivo experiments showed that the Venetoclax combined with Ixazomib group could effectively reduce the in vivo(4)transcriptome sequencing showed that NF-κB-related pathways were altered after the combination of Venetoclax and Ixazomib,and WB further verified that NF-κB protein expression was reduced in the combination group compared with the single drug group.Conclusion In in vivo experiments,the combination of the two drugs was found to inhibit tumor growth and prolong survival in AML xenograft mice by constructing an animal model;transcriptome sequencing technology was used to analyze the differential expression of the two drugs in the combination of Venetoclax and Ixazomib relative to the single drug group and the control group.The sequencing results were verified by Western blot after enrichment of the signaling pathway,and the synergistic mechanism of the two drugs was mainly in the NF-κB signaling pathway.The combination of Venetoclax and izatzomib can effectively kill AML cells,and the combination of the two drugs has the potential to be further investigated as a new clinical treatment option for AML. |
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